Rabbit Anti-Goat IgG H&L (HRP) (ab6741)

Submit a review Q&A (20)References (102)

Overview

  • Product name

    Rabbit Anti-Goat IgG H&L (HRP)
    See all IgG secondary antibodies

  • Host species

    Rabbit

  • Target species

    Goat

  • Tested applications

    Suitable for: Dot blot, ELISA, IHC-P, IHC-Fr, Immunomicroscopy, ICC/IF, WBmore details

  • Immunogen

    Goat IgG whole molecule

  • Conjugation

    HRP

Properties

  • Form

    Liquid

  • Storage instructions

    Shipped at 4°C. Store at +4°C.

  • Storage buffer

    pH: 7.20
    Preservative: 0.01% Gentamicin sulphate
    Constituents: 0.42% Potassium phosphate, 0.87% Sodium chloride, 1% BSA
  • Concentration information loading...

  • Purity

    Affinity purified

  • Purification notes

    This product was prepared from monospecific antiserum by immunoaffinity chromatography using Goat IgG coupled to agarose beads followed by solid phase adsorption(s) to remove any unwanted reactivities.

  • Conjugation notes

    Horseradish Peroxidase (HRP)

  • Clonality

    Polyclonal

  • Isotype

    IgG

  • Research areas

Applications

Our Abpromise guarantee covers the use of ab6741 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Dot blot Use at an assay dependent concentration.
ELISA 1/50000 - 1/100000.
IHC-P Use at an assay dependent concentration.
IHC-Fr Use at an assay dependent concentration.
Immunomicroscopy Use at an assay dependent concentration.
ICC/IF 1/1000 - 1/5000.
WB 1/5000 - 1/20000. PubMed: 17369188

Images

  • Western blot - Rabbit Anti-Goat IgG H&L (HRP) (ab6741)
    Western blot - Rabbit Anti-Goat IgG H&L (HRP) (ab6741)
    Ab6741 was used at dilution 1/5000 with the primary antibody ab23938 in WB. See the review on ab23938.
  • ELISA - Rabbit Anti-Goat IgG H&L (HRP) (ab6741)
    ELISA - Rabbit Anti-Goat IgG H&L (HRP) (ab6741)This image is courtesy of an anonymous Abreview.
    Ab6741 was used at dilution 1/1000 with the primary antibody ab2247 in ELISA. See the review on ab2247.

References

This product has been referenced in:

  • Li J  et al. The m6A demethylase FTO promotes the growth of lung cancer cells by regulating the m6A level of USP7 mRNA. Biochem Biophys Res Commun 512:479-485 (2019). Read more (PubMed: 30905413) »
  • Fostok S  et al. Connexin 43 Loss Triggers Cell Cycle Entry and Invasion in Non-Neoplastic Breast Epithelium: A Role for Noncanonical Wnt Signaling. Cancers (Basel) 11:N/A (2019). Read more (PubMed: 30857262) »
See all 102 Publications for this product

Publishing research using ab6741? Please let us know so that we can cite the reference in this datasheet.

Customer reviews and Q&As

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1-10 of 20 Abreviews or Q&A

Question

DO you know the expirate date of conjugated secondary antibodies? The secondary antibodies are same that primaries (4°C 1-2 Weeks and then to -20°C) https://www.abcam.com/Rabbit-polyclonal-Secondary-Antibody-to-Goat-IgG-HL-HRP-ab6741.html

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Answer

We have recently concluded a stability study for conjugated antibodies. Based on the results of this study we have decided to issue the following guidelines: Biotin conjugates +4°C short term. Aliquot and store at -20°C. Avoid freeze/thaw cycles. Alkaline phosphatase conjugates +4°C short term. Aliquot and store at -20°C. Avoid freeze/thaw cycles. HRP conjugates +4°C. Do not freeze. FITC conjugates +4°C. Do not freeze. In general an antibody stored at +4C may have a lifespan of about 1 year.

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Question

I have purchased the following antibodies from you. Could you please send me Material Safety Data Sheet (MSDS) for these items?
1. Anti-beta Actin antibody (Cat: ab8229)
2. Rabbit polyclonal Secondary Antibody to Goat IgG - H&L (HRP) (Cat: ab6741)

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Answer

Thank you for contacting us.

Here are the links for the MSDS for the 2 products:

For ab6741: https://www.abcam.com/ps/Products/6/ab6741/Msds/ab6741_WPNA_EN.pdf
For ab8229: https://www.abcam.com/ps/Products/8/ab8229/Msds/ab8229_WPNA_EN.pdf

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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Question

What is best secondary antibody you can offer for ab10256.

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Answer

Thank you for contacting us.

Please check the datasheet and selct on basis of conc, price and volume. The compatible products are ab102345, ab102354, ab5755, ab97100, ab97105 and ab6741.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Use our products? Submit an Abreview. Earn rewards!
https://www.abcam.com/abreviews

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Question

Our customer purchased 1 vial of ab6741 and 1 vial of ab6721 last December when we had some promotions.
He stored it 4C and openend it this morning to test.
But there were severe precipitation in the vial (ab6721, Lot.GR27131-6, ON.1005934).
I recommended to use after spin down but,it alreadyoccupied much portion of the vial,
he doesn't want to use it because hethinks he lost the volume about what he purchased 500ul of ab6721.
Please arrange a replacement vial for our customer.
Best regards,

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Answer

Thank you for your email. I am sorry to hear that you are experiencing problems with these antibodies.

The brown precipitates with HRP conjugated antibodies are very common. We recommend customers to pipette up and down and vortexing the vials which actually helps in dissolving the precipitates. As a final measure we also recommend centrifuging out the precipitates. It is true that it will affect the volume however because the product is at high conc. the effect will not be huge, only few mictolitres will be lost.

Please do not hesitate to use the antibodies in applications as recommended on the datasheet.

Please also note that we recommend storing this product at 4C for 1-2 weeks only, then it should be stored at -20C. The customer have stored the antibody at 4C since December 2011 which actually voids the Abpromise guarantee. Unfortunately replacement vials or refunds cannot be offered. However we are happy to troubleshoot he problem in case these products does not work.

I hope this information will be helpful. Should you have any question please do not hesitate to contact me.

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Question



Thanks for your reply. I did get some non-specific binding with the secondary only control but not as high as with both primary and secondary. I am using a very sensitive ECL reagent that we are developing and I believe I get the background in the secondary only control because I am doing long exposures to try and see the specific signal.

I would appreciate a replacement for the Primary antibody and I thank you for your help in resolving the matter.

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Answer

Thank you for your reply.

I was wondering if ab8227 would be a good replacement for you:

https://www.abcam.com/beta-Actin-antibody-Loading-Control-ab8227.html



If it is, then please let me know and I will get it sent out to you as soon as possible.

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Question



I recieved ab7125 as a replacement for ab6741 and tried it. I still have the same problem of non-specific background binding. I still believe as I originally thought that the primary antibody ab8229 may not be working. I have run controls where I use purified actin (from Sigma) and even though I can see some bands, the signal to noise ratio and detection limit indicate that this is from non-specific binding of the secondary antibody. I can send you a scan of the film if you would like.

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Answer

Thank you for your reply.

I am sorry the replacement secondary did not solve this problem. Do you get the same non-specific binding pattern when you do your secondary only control? I ask, because if you do, then that does indicate that the issue is with the secondary, not the primary (as was the case with the first secondary antibody you received).

If however, you are not seeing the non-specific binding in your secondary only controls, then I agree that the primary antibody is the problem and therefore I would be happy to send another one free of charge to resolve the matter.

I look forward to your replay and helping to resolve this issue.

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Question

I would be happy to try the replacement.

Thanks.

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Answer

Thank you for your reply.

As ab6741 did not prove to be successful for you, I am sending you ab7125 as a free of charge replacement. Your ne order number is ******* and you should have the new product by 10.30am tomorrow morning.

If there is anything else I can help you with, please let me know.

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Question

Regarding product ab6741, I did change the blocking reagent to BSA with no success. There was still high background. I also tried using blocking agent at every step, i.e. I put the blocking agent in with the secondary antibody. In that case I got no signal.

The antibody ab6741 was ordered on*******with PO number *******.

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Answer

Thank you for your reply.

I am sorry that you are continuing to have problems with ab6741 in western blot. Therefore I would like to send you ab7125 as a free of charge replacement:

https://www.abcam.com/Donkey-polyclonal-Secondary-Antibody-to-Goat-IgG-HL-HRP-pre-adsorbed-ab7125.html

Please let me know if this is ok with you, or if you would prefer a refund and I will be happy to have it sent out as soon as possible.

Looking forward to your reply.

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Question

High background from the secondary antibody.

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Answer

Thank you for contacting Abcam earlier today.

If you are still experiencing any issues with ab6741 in western blot after trying the protocol tips that we discussed, please let me know and I will be happy to look into sending a new antibody to try out.

Please let me know if there is anything else I can help you with.

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Question

I'm confused because when I had inquiry about the precipitation before, I received emailusing them with gentle vortex but,
this time, you answered spin down it and use only supernatant.
Let me know which method is right.


Best regards,

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Answer

Thank you for your reply.

I can confirm that for the HRP conjugated antibodies the approach that Irecommended is the correct one. Spining the vial down will prevent free HRP to bind to un-specifically to samples and therefore avoid increased background.

If the antibody is not purified and therefore also not conjugated then we would recommend gentle vortexing. This could be whole antiserum for example.
In this casethe antibody does contain proteins and other solutes it would not be abnormal to see a little cloudiness or precipitation, especially after being stored at a low temperature and/or for a long period of time. Most of the time bringing the antibody up to room temperature, mixing it well and then diluting it to a working concentration will help incorporate any protein aggregates and precipitation.

I hope this information will explain the different suggestions we gave and I apologise for the confusion.

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